Isolation, production and optimization of endogenous alkaline protease from in-situ sludge and its evaluation as sludge hydrolysis enhancer.

Abstract

Bioconversion (e.g. anaerobic fermentation and compost) is the common recycling method of waste activated sludge (WAS) and its hydrolysis, as the rate-limiting step of fermentation, could be accelerated by protease. However, the commercial protease was unstable in a sludge environment, which increased the cost. An endogenous alkaline protease stable in sludge environment was screened in this study and its suitability for treating the sludge was analyzed. The optimal production medium was determined by Response Surface Methodology as starch 20 g/L, KH2PO4 4 g/L, MgSO4·7H2O 1 g/L, sodium carboxy-methyl-cellulose 4 g/L, casein 4 g/L and initial pH 11.3, which elevated the yield of protease by up to 15 times (713.46 U/mL) compared with the basal medium. The obtained protease was active and stable at 35 °C-50 °C and pH 7.0-11.0. Furthermore, it was highly tolerant to sludge environment and maintained high efficiency of sludge hydrolysis for a long time. Thus, the obtained protease significantly hydrolyzed WAS and improved its bioavailability. Overall, this work provided a new insight for enzymatic treatment of WAS by isolating the endogenous and stable protease in a sludge environment, which would promote the resource utilization of WAS by further bioconversion.